Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
Journal of Life Science
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v.25
no.1
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pp.53-61
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2015
In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.
This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.
Fe2O3 nanoparticles with the mixed structure of cubic and nanorod were synthesized by precipitation, hydrothermal, sol-gel method, etching process and heat treatment. Fe2O3/TiO2 core-shell (CS) of type Fe2O3@TiO2 composite was fabricated on a 20 nm nanolayer of TiO2 coated on the surface of Fe2O3 nanoparticles. Fe2O3/TiO2 yolk-shell (YS) composite was prepared by chemical etching and heat treatment of Fe2O3/TiO2 CS nanoparticles. Physical properties of Fe2O3, Fe2O3@TiO2 CS and Fe2O3@TiO2 YS nanoparticles were characterized by FE-SEM, HR-TEM and X-ray diffraction. The solar reflectance, commission internationale de l'Elcairage (CIE) color coordinate and heat shield temperatures of Fe2O3, CS and YS type Fe2O3@TiO2 pigments filled with poly acrylate (PA) paints were investigated by UV-Vis-NIR spectrometer and homemade heat shield temperature measuring device. The Fe2O3@TiO2 YS red pigment filled PA composite exhibited excellent near infrared light reflecting performance and also reduced the heat shield temperature of 13 ℃ than that of Fe2O3 filled counterparts.
Fluxapyroxad is classified as carboxamide fungicide that inhibits succinate dehydrogenase in complex II of mitochondrial respiratory chain, which results in inhibition of mycelial growth within the fungus target species. This study was carried out to assure the safety of fluxapyroxad residues in agricultural products by developing an official analytical method. A new, reliable analytical method was developed and validated using High Performance liquid Chromatograph-UV/visible detector (HPLC-UVD) for the determination of fluxapyroxad residues. The fluxapyroxad residues in samples were extracted with acetonitrile, partitioned with dichloromethane, and then purified with silica solid phase extraction (SPE) cartridge. Correlation coefficient($R^2$) of fluxapyroxad standard solution was 0.9999. The method was validated using apple, pear, peanut, pepper, hulled rice, potato, and soybean spiked with fluxapyroxad at 0.05 and 0.5 mg/kg. Average recoveries were 80.6~114.0% with relative standard deviation less than 10%, and limit of detection (LOD) and limit of quantification (LOQ) were 0.01 and 0.05 mg/kg, respectively. All validation parameters were followed with Codex guideline (CAC/GL 40). LC-MS (Liquid Chromatograph-Mass Spectrometer) was also applied to confirm the analytical method. Base on these results, this method was found to be appropriate fluxapyroxad residue determination and can be used as the official method of analysis.
Brown color characteristics and antioxidizing activities were investigated for Doenjang under different processing conditions. Doenjang A was prepared directly from Meju and saline solution whereas Doenjang B was Prepared after separating soy sauce by soaking for 45 days. Both Doenjangs were aged for up to 180 days. Antioxidizing activity was studied in relation to the brown color characteristics using fat-soluble extract and water-soluble extract of Doenjang. The intensity of brown color was higher in the water-soluble Doenjang extract than the fat-soluble Doenjang extract. In the UV-VIS scanning spectra, water-soluble Doenjang extracts showed significant changes as the aging proceeded, but fat-soluble Doenjang extract did not. Antioxidizing activity of fat-soluble Doenjang extract increased as the aging period extended; however, no significant difference was detected in the water-soluble extract. Overall, Doenjang A showed higher contents of amino acids, reducing sugar, brown color, and antioxidizing activity, and the antioxidizing ability was higher in water-soluble Doenjang extract rather than in the fat-soluble Doenjang extract.
Purpose: Ultraviolet (UV)-induced oxidative stress contributes to several adverse biological effects on skin. Many phenolic phytochemicals have been shown to have antioxidant properties and protect skin cells from UV-induced oxidative damage. In this study, we investigated whether or not Aralia elata (AE) has a protective effect against UVB-induced reactive oxygen species (ROS), ultimately leading to photoaging. Methods: Phenolic content of dried AE and antioxidant properties of AE extract in 70% ethanol weredetermined by measuring DPPH and ABTS radical scavenging activities and ferric reducing antioxidant power (FRAP). The effect of AE extract on cellular ROS generation and expression levels of oxidative stress-response proteins such as superoxide dismutase (SOD)-1, catalase, nuclear factor-erythroid 2-related factor (Nrf)-2, and heme oxygenase (HO)-1 in UVB-irradiated ($75mJ/cm^2$) human keratinocytes (HaCaT) were further determined by 2'-7'-dichlorofluoresceine diacetate assay and Western blotting, respectively. Results: The total phenolic and flavonoid contents of dried AE were 20.15 mg tannic acid/g and 18.75 mg rutin/g, respectively. The $IC_{50}$ of AE extract against DPPH radical was $98.5{\mu}g/mL$, and ABTS radical scavenging activity and FRAP upon treatment with $1,000{\mu}g/mL$ of AE extract were $41.8{\mu}g\;ascorbic\;acid\;(AA)\;eq./mL$ and $29.7{\mu}g\;AA\;eq./mL$,m respectively. Pretreatment with AE extract significantly reduced (p < 0.05) ROS generation compared to that in UVB-irradiated control HaCaT cells. Pretreatment with AE extract reversed reduction of Nrf-2 and SOD-1 protein expression and induction of HO-1 protein expression caused by UVB exposure in HaCaT cells, whereas it did not affect catalase expression. Conclusion: AE extract in 70% ethanol demonstrated a protective effect against UVB-induced oxidative stress and decreased expression of Nrf-2 and SOD-1 in human keratinocytes. These findings suggest that AE ethanol extract might have potential as a natural resource for a skin anti-photoaging product in the food and cosmetic industry.
Specific carotenoids and astaxanthin biosynthesis power of Phaffia rhodozyma mutant 876, which was obtained after NTG a and UV treatments, was higher than those of the wild type by 40% and 50%, respectively. The mutant strain did not show t the catabolite repression even at 22% (w/v) glucose concentration. The optimum C{N ratio was 2.0, and the optimum t temperature and initial pH were $22^{\circ}C$ and 6.0, respectively. 80th cell growth and astaxanthin formation decreased drastically a as the fermentation temperature was increased over $22^{\circ}C$, whereas they were comparable in the pH range between 5.0 and 7 7.0. Inoculum size did not affect the final cell density nor the carotenoids biosynthesis, and 3%(v/v) was selected as optimal. H Higher dissolved oxygen concentration facilitated astaxanthin biosynthesis, and aeration rate of 1.0 v/0/m and agitation speed of 400 rpm were selected as optimum. The final cell dens때 of 43.3 g/L and the volumetric astaxanthin and carotenoids concentrations of 110.6 mg/L and 149.4 mg/L, respectively, were obtained. The specific carotenoids concentration was 3.45 m mg{g-yeast(dry). Yx/s and Yp/s values of 0.37 and 1.08 were obtained. The result of this study will provide basic information u useful for mass production of astaxanthin from P. rhodozyma fermentation.
Journal of the Korea Academia-Industrial cooperation Society
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v.19
no.9
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pp.279-287
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2018
Sealants are an important element of modern architecture and serve as a building protection against weathering by providing barriers against ingress of moisture, air, and other materials. Exposure to a variety of environments often reduces lifespan due to changes in physical, chemical and mechanical characteristics, and UV, humidity, and temperature expansion are important issues that are directly related to durability. In this study, a combined deterioration test chamber was developed to simulate the environment of the open air as an instrument for verifying the durability of structural sealing materials indoors. In order to replicate special weather conditions, such as yellow dust, acid rain, and contamination by microorganisms, it was deemed impossible to replicate the outdoor environment by 100 %, and the results of the results of the results of the external exposure test of the structural sealant and the combined deterioration testing device. As a result of the displacement test of the outdoor exposure test, it was determined that the sealant was breaking apart and that it would be smooth, and the displacement would be up to three times greater than the initial material value of 1 year. The displacement test results of the combined deterioration test device show the tendency to deteriorate, decreasing the elasticity and tensile characteristics. In the case of denatured silicon, the current 400 cycles have been completed to confirm 12 months of degradation of the external exposure. The deformation of the test specimen cannot be verified with the naked eye, so it is considered that the conditions of the specimen are more stable than the silicon sealant. As a result of the outdoor exposure test, if the combined deterioration test device is structured and proposed in the relevant guidance or specification, the anticipated lifespan of 12 months in the actual use environment can be verified indoors and below 3 months later, economically.
An experiment was conducted to investigate the dietary effects of a transgenic Aspergillus oryzae(AO) culture on the performance, egg quality and intestinal microflora of layers. A total of 840 Hy-line Brown layers of 39wks old were assigned to one of the following 7 dietary treatments: control(C), C+0.2% AO culture, C+0.5% AO culture, C+0.2% transgenic AO culture, C+0.5% transgenic AO culture, C+0.2% transgenic mutant AO culture, and C+0.5% transgenic mutant AO culture. The transgenic AO was made by inserting Salmonella gallinarum gene to AO. And the transgenic mutant AO was made by inserting Salmonella gallinarum gene to mutant AO which was mutated by UV irradiation. Each treatment was replicated six times with 20 birds housed in 2 bird cage. Twenty birds units were arranged according to completely randomized block design. Feeding trial lasted for 8wks under 16 hour lighting regimen. Laying performance and egg quality were significantly(P<0.05) affected by the treatments. Transgenic AO culture supplementation at the level of 0.2% significantly increased egg production, while its egg weight was significantly decreased compared to that of the control. Feed intake and feed conversion ratio(FCR) were not significantly different among the AO treatments and the control. The eggshell strength of the AO treatments was significantly higher than that of the control. Transgenic mutant AO culture supplemented at the level of 0.5% significantly increased egg yolk color. Intestinal microflora were significantly(P<0.05) affected by the treatments. The cfu of Lactobacilli spp. significantly increased and those of Salmonella species and E. coli decreased in the AO treatments. The transgenic AO and transgenic mutant AO culture were more effective than the AO culture in reducing the cfu of Salmonella species and E. coli. It is concluded that supplementation of the transgenic AO culture at the level of 0.2% could be recommended for the improvement of egg production. Supplementation of transgenic AO or transgenic mutant AO culture at 0.2% level effectively controlled intestinal Salmonella species population.
Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.
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